Fig 3.

Antibiotic-treated rats treated with gut microbiota transfer showed similar anxiety-like behaviours. (a) Illustration of experimental design. (b) Feeding mice antibiotics reduced alpha diversity of microbiota (P<0.05). (c) Feeding mice antibiotics reduced bacterial load (P<0.05) by semiquantitative real-time polymerase chain reaction (PCR). (d and e) Quantification of Firmicutes spp. and Verrucomicrobia spp. by quantitative PCR in 16s rRNA extracted from faeces after faecal microbiota transfer (FMT) (n=6 per group, ∗P<0.05 vs Control group, ^†P<0.05 vs FMT-Con group). (f and g) Levels of 5-HT and 5-HIAA were increased in the FMT-Multi group compared with the FMT-Con group (P<0.05). (h–j) Open Field Test (OF test) of FMT-Con, FMT-Multi, FMT-Multi+Vehicle, and FMT-Multi+ZIP groups. Compared with the FMT-Con group and the FMT-Multi+ZIP group, the FMT-Multi and FMT-Multi+Vehicle groups exhibited markedly less time in the centre (i, P<0.05). No statistical differences were found between groups in movement speed (h) and total distance (j, P>0.05). (k–m) Elevated plus maze test (EPM) of different groups. No statistical difference was found between groups in total distance (m, P>0.05), whereas the FMT-Multi and FMT-Multi+Vehicle groups showed a reduction in frequency (k) and time spent in the open arm (l, P<0.05). One-way analysis of variance followed by Tukey post hoc test. n=10 for each group, ∗P<0.05.5-HIAA, 5-hydroxyindoleacetic acid; 5-HT, 5-hydroxytryptamine; Abx, antibiotic mix in the drinking water for 2 weeks; Multiple AS, multiple sevoflurane/surgery exposure; PND, postnatal day; ZIP, ziprasidone 2.5 mg kg⁻¹ i. p. daily for 7 days.